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Effects of RU486 and indomethacin on meiotic maturation, formation of extracellular matrix, and progesterone production by porcine oocyte-cumulus complexes
- Source :
- Domestic Animal Endocrinology. 48:7-14
- Publication Year :
- 2014
- Publisher :
- Elsevier BV, 2014.
-
Abstract
- This study was designed to determine whether inhibition of either cyclooxygenase-2 (COX-2) by indomethacin or progesterone receptor (PR) by PR antagonist, RU486, affects oocyte maturation, progesterone production, and covalent binding between hyaluronan (HA) and heavy chains of inter-α trypsin inhibitor, as well as expression of cumulus expansion-associated proteins (HA-binding protein, tumor necrosis factor α-induced protein 6, pentraxin 3) in oocyte-cumulus complexes (OCCs). The experiments were based on freshly isolated porcine OCC cultures in which the consequences of PR and COX-2 inhibition on the final processes of oocyte maturation were determined. Granulosa cells (GCs) and OCCs were cultured in medium supplemented with FSH/LH (both 100 ng/mL) in the presence/absence of RU486 or indomethacin. Western blot analysis, (3)H-glucosamine hydrochloride assay, immunofluorescence, and radioimmunoassay were performed. Only treatment with RU486 (25 μM) caused a decrease in the number of oocytes that reached germinal vesicle breakdown and metaphase II stage compared with indomethacin (100 μM) or FSH/LH treatment alone after 44 h. All treated OCCs synthesized an almost equal amount of HA. Heavy chains (of inter-α trypsin inhibitor)-HA covalent complexes were formed during in vitro FSH/LH-stimulated expansion in RU486- or indomethacin-treated OCCs. Follicle-stimulating hormone/LH-induced progesterone production by OCCs was increased in the presence of RU486 after 44 h. In contrast, a decrease of FSH/LH-stimulated progesterone production by GCs was detected in the presence of either RU486 or indomethacin after 72 h. We suggest that the PR-dependent pathway may be involved in the regulation of oocyte maturation. Both PR and COX-2 regulate FSH/LH-stimulated progesterone production by OCCs and GCs.
- Subjects :
- endocrine system
medicine.medical_specialty
Swine
Trypsin inhibitor
Indomethacin
Biology
Hormone Antagonists
Endocrinology
Food Animals
Western blot
Internal medicine
Progesterone receptor
medicine
Animals
Cyclooxygenase Inhibitors
Hyaluronic Acid
Progesterone
Cumulus Cells
Germinal vesicle
medicine.diagnostic_test
Radioimmunoassay
Luteinizing Hormone
Oocyte
In vitro
Extracellular Matrix
In Vitro Oocyte Maturation Techniques
Mifepristone
Serum Amyloid P-Component
C-Reactive Protein
medicine.anatomical_structure
Gene Expression Regulation
Oocytes
Animal Science and Zoology
Tumor necrosis factor alpha
Follicle Stimulating Hormone
Carrier Proteins
Cell Adhesion Molecules
hormones, hormone substitutes, and hormone antagonists
Subjects
Details
- ISSN :
- 07397240
- Volume :
- 48
- Database :
- OpenAIRE
- Journal :
- Domestic Animal Endocrinology
- Accession number :
- edsair.doi.dedup.....31c665573a952f2f088f78bbe7a50fd8